Literature summary for 2.7.1.146 extracted from

Castro-Fernandez, V.; Herrera-Morande, A.; Zamora, R.; Merino, F.; Gonzalez-Ordenes, F.; Padilla-Salinas, F.; Pereira, H.M.; Brandao-Neto, J.; Garratt, R.C.; Guixe, V.
Reconstructed ancestral enzymes reveal that negative selection drove the evolution of substrate specificity in ADP-dependent kinases (2017), J. Biol. Chem., 292, 15598-15610 .

Search for more literature for 2.7.1.146

Cloned(Commentary)

Cloned (Comment)	Organism
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Pyrococcus horikoshii
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Methanocaldococcus jannaschii
phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	uncultured bacterium

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme mutant E72A, hanging drop vapor diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 25 mM HEPES/NaOH, pH 7.8, 30 mM MgCl2, 20 mM fructose 6-phosphate, and 20 mM AMP with 0.002 ml of reservoir solution containing 18% PEG 3350 and 0.15 M NaI, 3 days at 18°C, X-ray diffraction structure determination and analysis at 2.61 A resolution, molecular replacement using the structure of Pyrococcus horikoshii ADP-PFK (PDB ID 3DRW) split into small and large domains as search models	uncultured bacterium

Crystallization (Comment)

Organism

purified enzyme mutant E72A, hanging drop vapor diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 25 mM HEPES/NaOH, pH 7.8, 30 mM MgCl2, 20 mM fructose 6-phosphate, and 20 mM AMP with 0.002 ml of reservoir solution containing 18% PEG 3350 and 0.15 M NaI, 3 days at 18°C, X-ray diffraction structure determination and analysis at 2.61 A resolution, molecular replacement using the structure of Pyrococcus horikoshii ADP-PFK (PDB ID 3DRW) split into small and large domains as search models

uncultured bacterium

Protein Variants

Protein Variants	Comment	Organism
E72A	site-directed mutagenesis, the mutant shows reduced glucokinase and phosphofructokinase activities compared to wild-type enzyme	uncultured bacterium

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	enzyme PhPFK is not significantly inhibited by D-glucose	Pyrococcus horikoshii

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0063	-	D-fructose 6-phosphate	pH 6.5, 40°C	Methanocaldococcus jannaschii
0.056	-	D-fructose 6-phosphate	pH 6.5, 40°C	Pyrococcus horikoshii
0.119	-	D-fructose 6-phosphate	pH 6.5, 40°C, wild-type enzyme	uncultured bacterium
2.8	-	D-fructose 6-phosphate	pH 6.5, 40°C, mutant E72A	uncultured bacterium

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pyrococcus horikoshii
Mg2+	required	Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
ADP + D-fructose 6-phosphate	uncultured bacterium	-	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	Pyrococcus horikoshii	-	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	Methanocaldococcus jannaschii	-	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	Methanocaldococcus jannaschii ATCC 43067	-	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	Pyrococcus horikoshii ATCC 700860	-	AMP + D-fructose 1,6-bisphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	Q58999	-	-
Pyrococcus horikoshii	O59355	-	-
Pyrococcus horikoshii ATCC 700860	O59355	-	-
uncultured bacterium	-	synthetic construct	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	uncultured bacterium
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	Pyrococcus horikoshii
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ADP + D-fructose 6-phosphate	-	uncultured bacterium	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	-	Pyrococcus horikoshii	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	-	Methanocaldococcus jannaschii	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	-	Methanocaldococcus jannaschii ATCC 43067	AMP + D-fructose 1,6-bisphosphate	-	?
ADP + D-fructose 6-phosphate	-	Pyrococcus horikoshii ATCC 700860	AMP + D-fructose 1,6-bisphosphate	-	?
additional information	the enzyme also shows glucokinase activity, a bifunctional AncPFK/GK ancestor enzyme	uncultured bacterium	?	-	-
additional information	the enzyme from Methanococcus jannaschii also shows glucokinase activity, a bifunctional MjPFK/GK	Methanocaldococcus jannaschii	?	-	-
additional information	the PFK from Pyrococcus horikoshii (PhPFK) has no activity with glucose	Pyrococcus horikoshii	?	-	-
additional information	the enzyme from Methanococcus jannaschii also shows glucokinase activity, a bifunctional MjPFK/GK	Methanocaldococcus jannaschii ATCC 43067	?	-	-
additional information	the PFK from Pyrococcus horikoshii (PhPFK) has no activity with glucose	Pyrococcus horikoshii ATCC 700860	?	-	-

Synonyms

Synonyms	Comment	Organism
ADP-dependent glucokinase	-	uncultured bacterium
ADP-GK	-	uncultured bacterium
ancGK/PFK	-	uncultured bacterium
MjPFK/GK	-	Methanocaldococcus jannaschii
More	see also EC 2.7.1.147	uncultured bacterium
More	see also EC 2.7.1.147	Methanocaldococcus jannaschii
PFK	-	Pyrococcus horikoshii
PhPFK	-	Pyrococcus horikoshii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
40	-	assay at	uncultured bacterium
40	-	assay at	Pyrococcus horikoshii
40	-	assay at	Methanocaldococcus jannaschii

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.33	-	D-fructose 6-phosphate	pH 6.5, 40°C, wild-type enzyme	uncultured bacterium
0.38	-	D-fructose 6-phosphate	pH 6.5, 40°C, mutant E72A	uncultured bacterium
2.7	-	D-fructose 6-phosphate	pH 6.5, 40°C	Methanocaldococcus jannaschii
21	-	D-fructose 6-phosphate	pH 6.5, 40°C	Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	-	assay at	uncultured bacterium
6.5	-	assay at	Pyrococcus horikoshii
6.5	-	assay at	Methanocaldococcus jannaschii

Cofactor

Cofactor	Comment	Organism	Structure
ADP	-	Pyrococcus horikoshii
ADP	-	Methanocaldococcus jannaschii

General Information

General Information	Comment	Organism
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	uncultured bacterium
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	Pyrococcus horikoshii
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	Methanocaldococcus jannaschii
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	uncultured bacterium
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	Pyrococcus horikoshii
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	Methanocaldococcus jannaschii

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.136	-	D-fructose 6-phosphate	pH 6.5, 40°C, mutant E72A	uncultured bacterium
2.77	-	D-fructose 6-phosphate	pH 6.5, 40°C, wild-type enzyme	uncultured bacterium
375	-	D-fructose 6-phosphate	pH 6.5, 40°C	Pyrococcus horikoshii
428.6	-	D-fructose 6-phosphate	pH 6.5, 40°C	Methanocaldococcus jannaschii